Registration Dossier

Administrative data

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Effects on fertility

Description of key information

There were no effects on fertility

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018-2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Specific details on test material used for the study:
Test item: Hydrogenated Rosin Alcohols.
Intended use: Industrial use.
Appearance: Colorless tacky resin.
Storage conditions: At ambient temperature (15 to 25°C), in the dark.
Supplier: Sponsor.
Batch number: VR-096
Expiry date: 08 January 2019 - One year from receipt (08 January 2018).
Purity: 100%
Supplier’s responsibilities: Characterization of the test item and the documentation of the methods of synthesis, fabrication or derivation and stability.
Archive sample: A 0.5 g representative sample was taken from each batch of test item. This sample was placed in a well closed glass container and stored in the archives under the same conditions as the bulk material.
Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
Strain/Species Crl:CD(SD) rat.
Supplier Charles River (UK) Ltd.
Number of animals ordered 44 males and 48 females.

Spare animals were removed from the study room after treatment commenced.

Duration of acclimatization Males: six days before commencement of treatment.
Females: 20 days before commencement of treatment.
Age of the animals at the start of treatment Males: 69 to 75 days old. Females: 83 to 89 days old.
Weight range of the animals at the start of treatment Males: 329 to 407 g. Females: 210 to 275 g.
Sex:
male/female
Details on test animals or test system and environmental conditions:
Animal Care and Husbandry
Environmental Control

Rodent facility Limited barrier - to minimize entry of external biological and chemical agents and to minimize the transference of such agents between rooms.
Air supply F iltered fresh air which was passed to atmosphere and not recirculated.
Temperature and relative humidity Monitored and maintained within the range of 20-24ºC and 40-70%. Although conditions were occasionally outside the indicated ranges, these deviations were minor and/or of short duration and were not considered to have influenced the health of the animals and/or the outcome of the study.
Lighting Artificial lighting, 12 hours light : 12 hours dark.
Electricity supply Public supply with automatic stand-by generators.
Animal Accommodation
Cages Cages comprised of a polycarbonate body with a stainless steel mesh lid; changed at appropriate intervals. Solid (polycarbonate) bottom cages were used during the acclimatization, pre-pairing, treatment, gestation, littering and lactation periods. Grid bottomed polypropylene cages were used during pairing. These were suspended above absorbent paper which was changed daily during pairing.
Cage distribution The cages were distributed on the racking to equalize, as far as possible, environmental influences amongst the groups.
Bedding Solid bottom cages contained softwood based bark-free fiber bedding, which was changed at appropriate intervals each week.
Number of animals per cage Pre-pairing up to four animals of one sex, Pairing one male and one female , Males after mating up to four animals
Gestation one female
Lactation one female + litter
3.4.3 Environmental Enrichment
Aspen chew block A soft white untreated wood block; provided to each cage throughout the study (except during pairing and lactation) and replaced when necessary.
Plastic shelter Provided to each cage throughout the study (except during pairing and lactation) and replaced at the same time as the cages.
Nesting material Paper shavings were provided to each cage from Day 20 after mating and changed at the same frequency as the bedding.
3.4.4 Diet Supply
Diet SDS VRF1 Certified powdered diet.
A sample (250g) of each batch of diet used was retained within Pharmacy (frozen -10 to -30ºC) until finalization of the report. Samples were discarded after finalization of the report.
The diet contained no added antibiotic or other chemotherapeutic or prophylactic agent.
Availability Non-restricted (See Section 4).
3.4.5 Water Supply
Supply Potable water from the public supply via polycarbonate bottles with sipper tubes. Bottles were changed at appropriate intervals.
Availability Non-restricted.
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
Route Oral, via the diet.
Treated at Constant dietary concentrations (ppm) for each group.
Control (Group 1) Untreated diet of the same batch.
Frequency Continuously.
Diet A record of the usage of the diets was maintained on all occasions when food consumption was measured. This was performed using the initial weight of the diet container and an on-line data check on completion of the feeding procedure to ensure that all cages were fed the correct amount of diet. No significant discrepancy was found.
Details on mating procedure:
Pairing commenced After a minimum of two weeks of treatment.
Male/female ratio 1:1 from within the same treatment groups.
Duration of pairing Up to two weeks. of mating
Daily checks for evidence of mating Ejected copulation plugs in cage tray and sperm in the vaginal smear.
Day 0 of gestation When positive evidence of mating was detected.
Male/female separation Day when mating evidence was detected.
Pre-coital interval Calculated for each female as the time between first pairing and evidence of mating.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Stability and homogeneity: Before commencement of treatment, the suitability of the proposed mixing procedures was determined and specimen formulations at 100 and 20000 mg/mL were analyzed to assess the stability and homogeneity of the test item in the diet matrix.

Achieved concentration: Samples of each formulation prepared for administration in Week 1 and in the last week of treatment were analyzed for achieved concentration of the test item.
Duration of treatment / exposure:
Three groups of ten male and ten female rats received Hydrogenated Rosin Alcohols orally, via the diet, at concentrations of 4000, 7500 or 15000 ppm. Males were treated daily for two weeks before pairing, up to necropsy after 35 consecutive days. Females were treated daily for two weeks before pairing, throughout pairing, gestation and until Day 13 of lactation. Females were allowed to litter, rear their offspring and were killed on Day 13 of lactation
Frequency of treatment:
Daily
Details on study schedule:
Three groups of ten male and ten female rats received Hydrogenated Rosin Alcohols orally, via the diet, at concentrations of 4000, 7500 or 15000 ppm. Males were treated daily for two weeks before pairing, up to necropsy after 35 consecutive days. Females were treated daily for two weeks before pairing, throughout pairing, gestation and until Day 13 of lactation. Females were allowed to litter, rear their offspring and were killed on Day 13 of lactation
Dose / conc.:
4 000 ppm
Dose / conc.:
7 500 ppm
Dose / conc.:
15 000 ppm
No. of animals per sex per dose:
10
Control animals:
yes, concurrent no treatment
Positive control:
No
Parental animals: Observations and examinations:
Animals were inspected visually at least twice daily for evidence of ill-health or reaction to treatment. Cages were inspected daily for evidence of animal ill-health amongst the occupant(s). Any deviation from normal was recorded at the time in respect of nature and severity, date and time of onset, duration and progress of the observed condition, as appropriate.
During the acclimatization period, observations of the animals and their cages were recorded at least once per day.
Oestrous cyclicity (parental animals):
Dry and wet smears were taken as follows:
Dry smears For 15 days before pairing using cotton swabs.
Wet smears Using pipette lavage during the following phases:
For 14 days before treatment (all females including spares); animals that failed to exhibit 4-5 day cycles were not allocated to study.

After pairing until mating.

For four days before scheduled termination (nominally Days 10-13 of lactation).

Sperm parameters (parental animals):
Not measured
Litter observations:
Clinical observations Examined at approximately 24 hours after birth (Day 1 of age) and then daily thereafter for evidence of ill health or reaction to maternal treatment; these were on an individual offspring basis or for the litter as a whole, as appropriate.
Litter size Daily records were maintained of mortality and consequent changes in litter size from Days 1-13 of age.
Sex ratio of each litter Recorded on Days 1, 4, 7 and 13 of age.
Individual offspring body weights Days 1, 4, 7 and 13 of age.
Ano-genital distance Day 1 - all F1 offspring.
Nipple/areolae count Day 13 of age - male offspring.
Postmortem examinations (parental animals):
All adult animals were subject to a detailed necropsy. After a review of the history of each animal, a full macroscopic examination of the tissues was performed. All external features and orifices were examined visually. Any abnormality in the appearance or size of any organ and tissue (external and cut surface) was recorded and the required tissue samples preserved in appropriate fixative.
Postmortem examinations (offspring):
Premature deaths W here possible, a fresh macroscopic examination (external and internal) with an assessment of stomach for milk content was performed. Abnormal tissues were retained.
F1 offspring on Day 4 of age Blood sampling required (See Section 3.6.11).
Selected Day 4 offspring with no clinical observations discarded without examination.
Selected Day 4 offspring with clinical observations, examined externally, and retained pending possible future examination.
F1 offspring on Day 13 of age Blood sampling required (See Section 3.6.11).
All animals were subject to an external macroscopic examination; particular attention was paid to the external genitalia. Animals observed with external abnormalities were retained pending possible future examination
Thyroid glands were preserved from one male and one female in each litter.
Statistics:
Provided below as this section won't allow the full description to be included.
Reproductive indices:
Duration of gestation Time elapsing between the detection of mating and commencement of parturition.
Parturition observations From Day 20 after mating, females were inspected three times daily for evidence of parturition. The progress and completion of parturition was monitored, numbers of live and dead offspring were recorded and any difficulties observed were recorded.
Clinical signs:
no effects observed
Description (incidence and severity):
There were considered to be no signs seen that were related to treatment during detailed physical examination and arena observations for animals in the treatment, gestation or lactation period.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Overall body weight gains in males during treatment and females before pairing was low in males at 7500 ppm and females at 15000 ppm (73% and 70%, respectively); and, markedly low for males at 15000 ppm and females at 7500 ppm (61% and 52%, respectively).
During the first week of treatment, group mean body weight gain was markedly low at 15000 ppm (4 g and 1 g compared with 27g and 9g of the Control, for males and females respectively). Thereafter, there was a general improvement in body weight gains, however they remained low for males until Week 4.
Females receiving 15000 ppm commenced gestation with slightly low body weight, when compared to Control (94%), however during the first 2 weeks of gestation (Days 0 to 14) body weight gains were similar to Control. And, although the difference did not attain statistical significance, the overall body weight gain seen in females at 15000 ppm during gestation was slightly low and was primarily due to their performance during Gestation Days 14 to 20 (86% of Control). There was no effect on body weights during gestation for females at 4000 or 7500 ppm.
Body weight gain was unaffected by treatment during lactation. The group mean body weight value for females receiving 15000 ppm when they commenced lactation was slightly low when compared to Control.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food intake was lower than Control for males receiving 7500 or 15000 ppm throughout the treatment period, although statistical significance was only achieved in Weeks 1 (85% of Control at 15000 ppm) or 5 (90% of Control at 7500 ppm).
Prior to pairing, food intake of females receiving 15000 ppm was lower during Week 1 (84 % of Control) or in females at 7500 ppm in Week 2 (82% of Control).
During gestation, food intake of females receiving 7500 or 15000 ppm was lower during gestation days 0 to 14 (86 or 91% of Controls for period), but similar (100 or 96% of Controls for period) during gestation days 14 to 20.
During lactation, food intake of females receiving 15000 ppm was (90%) lower, although the differences did not attain statistical significance.
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The hematological investigation revealed a decreased hematocrit value (94%) in males receiving 15000 ppm although there were no corresponding changes in the other erythrocyte indices. In females, haematocrit concentration was also decreased in females receiving 7500 or 15000 ppm (89 or 87% of Control, respectively), with a consequential (since MCHC can be derived from hemaglobin and hematocrit values) increase in mean cell hemaglobin concentration (105 or 104% of Control, respectively), although the difference was offset since the hemaglobin concentration was also reduced (94 or 90%, of Control, respectively).
All differences from controls were minor, confined to one sex or lacked dose-response and were therefore attributed to normal biological variation. Such differences included the slightly low lymphocyte count in males at 15000 ppm (69% of Control), since there was no clear relationship to dose, was not present in females and may have been influenced by two particularly high values in the Control group. The lower basophil count in females at 15000 ppm (33% of Control) or the decreased prothrombin time in treated females (117, 114 or 118% at 4500, 7500 or 15000 ppm) since there was no relationship to dose and no clear effect seen in the males.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
There was a statistically significantly decrease of phosphorus in males at 15000 ppm and females at 4000, 7500 or 15000 ppm (77% and 56, 72 or 65%, respectively). There was an increase in alanine-amino transferase in males and females at 15000 ppm (141% and 115%, respectively), although statistical significance was only attained in males. There was also a decrease in triglyceride concentrations of males receiving 4000, 7500 or 15000 ppm (65, 58 or 64%, respectively), although this was not present in the females. Lastly, plasma cholesterol concentrations in females receiving 7500 or 15000 ppm were increased (130 or 129%, respectively).
All other differences from controls including those that attained statistical significance, were minor, confined to one sex or lacked dose-response and were therefore attributed to normal biological variation. Such differences included the decrease in bile acids in males (25% of Control), since no corresponding pattern was seen in females. And, the increased albumin (95% of Control) since the change was minor.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
The microscopic examination performed after 5 weeks of treatment revealed no test item related lesions. The incidence and distribution of all findings were considered to be unrelated to treatment.
Histopathological findings: neoplastic:
no effects observed
Other effects:
not specified
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
All females allocated to the study showed regular 4 or 5 day estrus cycles prior to the start of treatment. There was no effect of treatment on the stage of the estrus cycle during treatment or at termination.
Pre-coital interval, mating performance and fertility and gestation length were unaffected by treatment with Hydrogenated Rosin Alcohols. The gestation index was 100 % in all groups.
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
Pre-coital interval, mating performance and fertility and gestation length were unaffected by treatment with Hydrogenated Rosin Alcohols. The gestation index was 100 % in all groups.
There is one point of clarification needed for the F0-F1 in lactation. The report is clear that at the high dose, food consumption is decreased at the high dose compared to the control. The report is not as clear in the body text on the absolute food consumption and achieved dose along with pup body weights. Please see the data tables in the "Any other information on results including tables" section. During gestation, the achieved dose was unchanged over time. However, as expected, in lactation, the absolute achieved dose rose dramatically as a reflection of the increase in absolute food consumption. This is a normal finding in lactation for the dam to meet the nutritional needs of the pups. In this study, the laboratory did not adjust the dose to meet the increased food consumption, so across the board, the achieved doses were much higher than baseline, and in the high dose group, the achieved dose had doubled to 1944 mg/kg/day by lactation days 7-13.
In the OECD 422 study design, the high dose is set at either a limit dose of 1000 mg/kg/day or to the MTD if it is lower. In this study, the MTD and limit were essentially the same. So in the late days of gestation, the dams were exposed to a dose that was twice the intended high dose and twice the limit dose of the study. It is very clear that pup body weights decrease as maternal test article ingestion increases, and the effects seen on the offspring are a function of the gross overdose that the dams were receiving as opposed to a relevant dose that would be used to set a risk value.
Dose descriptor:
NOAEL
Effect level:
15 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Dose descriptor:
NOAEL
Effect level:
15 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reproductive function (oestrous cycle)
reproductive function (sperm measures)
reproductive performance
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
There was no second generation in the study design, so no parental generation either.
Clinical signs:
no effects observed
Description (incidence and severity):
There were no clinical signs observed among the F1 litters that were clearly attributable to parental treatment of the Test Item.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
There were slightly low implantation counts in the females receiving 7500 or 15000 ppm, compared to control (88 or 86%).
The total litter size was lower at 15000 ppm (89% of Control) as a result of the low implantation count, although the live litter size on Day 1, post implantation survival index, sex ratio and offspring survival indices were unaffected by treatment.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Mean offspring body weight on Day 1 of age were unaffected by treatment. However, overall body weight gain (Days 1 to 13 of age) was statistically significantly low in male or female offspring of parents receiving 15000 ppm (80 or 81% of Control, respectively). When examined closely (see data tables below) beginning around PND 4, dams began a drastic increse in food consumption (as expected) resulting in an acheived test artice dose that was almost two times the intake prior to parturition. This dose was approximately twice the limit dose in a DART study, and was twice the MTD, which was the top dose. So essentially the high dose dams were being grossly overdosed from day 4 of lactation, and that increase in dose past the limit and/or the MTD is reflected in the decrease in pup weights. This also explains why there was no weight decrease issues on PND 1, as the increase in food consumption had not started at that point.
Food consumption and compound intake (if feeding study):
not examined
Description (incidence and severity):
Pups were sacrificed before weaning, but see the above comments on body weight and the remarks and data tables on maternal food consumption and acheived test article dose.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Anogenital distance (AGD):
no effects observed
Description (incidence and severity):
The ano-gential distances of offspring from parents treated at 7500 or 15000 ppm was increased (107% for both male groups and 115 or 110% for females, respectively), when compared to Control. However, values were within the HCD range (2.9 to 4.8 mm for males and 1.8 to 2.6 mm for females) so no effect of treatment was inferred.
Nipple retention in male pups:
no effects observed
Description (incidence and severity):
No retained nipples were found
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no macroscopic findings observed in the offspring that died prior to scheduled termination or among those offspring killed on Day 13 of age that were attributable to parental treatment with the test item.
Histopathological findings:
not examined
Other effects:
not specified
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
There is one point of clarification needed for the F0-F1 in lactation. The report is clear that at the high dose, food consumption is decreased at the high dose compared to the control. The report is not as clear in the body text on the absolute food consumption and achieved dose along with pup body weights. Please see the data tables in the "Any other information on results including tables" section. During gestation, the achieved dose was unchanged over time. However, as expected, in lactation, the absolute achieved dose rose dramatically as a reflection of the increase in absolute food consumption. This is a normal finding in lactation for the dam to meet the nutritional needs of the pups. In this study, the laboratory did not adjust the dose to meet the increased food consumption, so across the board, the achieved doses were much higher than baseline, and in the high dose group, the achieved dose had doubled to 1944 mg/kg/day by lactation days 7-13.
In the OECD 422 study design, the high dose is set at either a limit dose of 1000 mg/kg/day or to the MTD if it is lower. In this study, the MTD and limit were essentially the same. So in the late days of gestation, the dams were exposed to a dose that was twice the intended high dose and twice the limit dose of the study. It is very clear that pup body weights decrease as maternal test article ingestion increases, and the effects seen on the offspring are a function of the gross overdose that the dams were receiving as opposed to a relevant dose that would be used to set a risk value.
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 944 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
Critical effects observed:
no
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Anogenital distance (AGD):
not specified
Nipple retention in male pups:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
An F2 generation was not bred, so this section is not populated
Reproductive effects observed:
no

Achieved Dose (mg/kg) in dams over Gestation and Lactation
Gestation Lactation
Group/Sex       Day Mean Day Mean
     0-7        7-14         14-20       0-20      1-4    4-7  7-13 1-13
2F 298 280 271 284 419 584 719 610
3F 518 469 508 498 745 1019 1086 984
4F 1058 1002 994 1019 1532 1952 1944 1843

Body Weight and Body Weight Change - Group Mean Values (g) for F1 Offspring
Group/Sex   Day of age (before bleed) Day of age (after bleed) Change
 1-4
change
 4-7
change
7-13 
change
1-13
    1 1@ 4 4 7 13        
1M Mean 6.9 6.9 9.9 9.9 15.1 27 2.9 5.2 11.9 201
  SD 0.64 0.64 1.18 1.18 1.73 3.47 0.74 0.61 2.14 3.09
  N 9 9 9 9 9 9 9 9 9 9
2M Mean 6.9 6.9 9.8 9.8 14.8 26.1 2.8 5.1 11.3 19.2
  SD 0.73 0.73 0.98 0.98 1.69 2.38 0.4 0.74 0.88 1.76
  N 10 10 10 10 10 10 10 10 10 10
3M Mean 6.9 7 9.7 9.7 14.9 26 2.8 5.2 11.1 19
  SD 0.92 0.82 1.3 1.3 1.94 3.37 0.54 0.73 1.76 2.8
  N 8 8 8 8 8 8 8 8 8 8
4M Mean 6.9 6.9 9.5 9.5 14 23** 2.6 4.5 9 16.1
  SD 0.35 0.37 0.72 0.72 0.54 0.82 0.5 0.27 0.44 0.57
  N 10 10 10 10 10 10 10 10 10 10

Body Weight and Body Weight Change - Group Mean Values (g) for F1 Offspring
Group/Sex   Day of age (before bleed) Day of age (after bleed) Change
 1-4
change
 4-7
change
7-13 
change
1-13
    1 1@ 4 4 7 13        
1F Mean 6.5 6.6 9.3 9.4 14.5 25.8 2.8 5.1 11.4 19.2
  SD 0.62 0.67 1.32 1.31 1.8 3.5 0.84 0.6 1.94 3.04
  N 9 9 9 9 9 9 9 9 9 9
2F Mean 6.4 6.4 9.3 9.3 14.1 25.3 2.8 4.8 11.2 18.8
  SD 0.67 0.68 1.01 1.02 1.87 2.67 0.53 0.93 0.88 2.16
  N 10 10 10 10 10 10 10 10 10 10
3F Mean 6.4 6.5 9.2 9.3 14.1 25.4 2.8 4.8 11.2 18.9
  SD 0.82 0.93 1.22 1.3 2.02 3.42 0.5 0.83 1.7 2.75
  N 8 8 8 8 8 8 8 8 8 8
4F Mean 6.4 6.4 8.9 9 13.3 21.9** 2.6 4.3* 8.6** 15.5**
  SD 0.3 0.25 0.63 0.61 0.42 0.77 0.4 0.34 0.57 0.65
  N 10 10 10 10 10 10 10 10 10 10
Conclusions:
Based on the results of this study it is concluded that the No-observed-adverse-effect-level (NOAEL) of Hydrogenated Rosin Alcohols for systemic toxicity is 15000 ppm (equivalent to 902 mg/kg/day for males and 1077 mg/kg/day for females) and for reproductive/developmental effects is 1944 mg/kg/day based on the reduced offspring body weight in lactation. This decrease is directly related to a gross overdose of the high dose females when the test article concentration was not adjusted in lactation to meet the increased food demands of the dam. This resulted in a maternal dose that was twice the limit dose of 1000 mg/kg/day and twice the MTD.
Executive summary:

The purpose of this study was the assessment of general systemic potential in rats, including a screen for reproductive/developmental effects and assessment of endocrine relevant endpoints, with administration of Hydrogenated Rosin Alcohols, for industrial use, by dietary administration for at least five weeks.

Three groups of ten male and ten female rats received Hydrogenated Rosin Alcohols orally, via the diet, at concentrations of 4000, 7500 or 15000 ppm. Males were treated daily for two weeks before pairing, up to necropsy after 35 consecutive days. Females were treated daily for two weeks before pairing, throughout pairing, gestation and until Day 13 of lactation. Females were allowed to litter, rear their offspring and were killed on Day 13 of lactation. The F1 generation received no direct administration of the test item; any exposure was in utero or via the milk. A similarly constituted Control group received the vehicle, untreated diet of the same batch.

During the study, clinical condition, detailed physical examination and arena observations, sensory reactivity observations, grip strength, motor activity, body weight, food consumption, hematology (peripheral blood), blood chemistry, thyroid hormone analysis, estrous cycles, pre-coital interval, mating performance, fertility, gestation length, organ weight and macroscopic pathology and histopathology investigations were undertaken.

The clinical condition, litter size and survival, sex ratio, body weight, ano-genital distance and macropathology for all offspring were also assessed. Nipple counts were performed on male offspring on Day 13 of age.

Mean achieved doses for males at 4000, 7500 or 15000 ppm were 240, 417 or 902 mg/kg/day, respectively. For females prior to pairing mean achieved doses were 283, 494 or 1077 mg/kg/day, respectively.

Mean achieved doses for females during gestation were 284, 498 and 1019 mg/kg/day, respectively, and for females during lactation were 610, 984 or 1843 mg/kg/day, respectively.

Analyses of samples for thyroxine (T4) obtained from Main study male animals and F1 offspring on Day 13 of age did not reveal any differences that could be attributed to treatment.

Administration of Hydrogenated Rosin Alcohols at dietary concentrations of 4000, 7500 or 15000 ppm had no adverse effect on clinical condition, sensory reactivity and grip strength, motor activity,hematology,estrous cycles, pre-coital interval, mating performance, fertility and gestation length, macropathology of the adults or micropathology.

For males during treatment, body weight gains and food intake were 73% or 61% of Controls, and for females prior to pairing body weights were 52% or 70% of Controls, at 7500 or 15000 ppm, respectively. Food intake remained low for females during gestation, at 7500 or 15000 ppm (86 or 91% of Controls during gestation days 0 to 14, respectively), and lactation, at 15000 ppm (90%), although reduced body weight gains, at 15000 ppm, was only seen in the later part of gestation (86% of Control from days 14 to 20).

Biochemical examination at the end of the treatment period revealed low phosphorus in males at 15000 ppm and in all groups of treated females (77% and 56, 72 or 65% at 4000, 7500 or 15000 ppm, respectively); an increase in alanine-amino transferase in males and females at 15000 ppm (141% and 115%, respectively); low triglyceride concentrations in treated males (65, 58 or 64% at 4000, 7500 or 15000 ppm, respectively); and, slightly high cholesterol concentrations in females receiving 7500 or 15000 ppm (130 or 129%, respectively). 

At scheduled termination, analysis of organ weights revealed high adjusted liver weights in all treated female groups(between 123 to 155%)and in males at 7500 or 15000 ppm(113 or 131%, respectively), with relationship to treatment.

Implantation counts were slightly low at 7500 or 15000 ppm (88 or 86%) but live litter size was unaffected. There was no effect of parental treatment on offspring clinical signs, sex ratio, offspring body weight on Day 1, survival, nipple counts, ano-genital distances or macropathology. 

Offspring body weight by Day 13 from parents treated at 15000 ppm was lower than Controls due to reduced growth at 19 to 20% lower than Controls.

Based on the results of this study it is concluded that the No-observed-adverse-effect-level (NOAEL) of Hydrogenated Rosin Alcohols for systemic toxicity and reproductive toxicity in the F0 is 15000 ppm (equivalent to 902 mg/kg/day for males and 1077 mg/kg/day for females) and for F1 reproductive/developmental effects is 1944 mg/kg/day based on the reduced offspring body weight. This decrease is directly related to a gross overdose of the high dose females when the test article concentration was not adjusted in lactation to meet the increased food demands of the dam.  This resulted in a maternal dose that was twice the limit dose of 1000 mg/kg/day and twice the MTD.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 077 mg/kg bw/day
Study duration:
subacute
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

In accordance with Annex IX, as there is no information suggesting further studies are necessary, no additional work is proposed.

Effects on developmental toxicity

Description of key information

No developmental toxicity was observed in an OECD 422 study at the limit dose/MTD. The was some fetal body weight loss around lactation day 14 due to the laboratory not reducing the concentration of the test article in the food to meet the inceasing consumption of the lactating dams. This resulted in acheived doses of ~ 2000 mg/kg/day, which is twice the limit dose and MTD.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 944 mg/kg bw/day
Study duration:
subacute
Species:
rat
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available

Justification for classification or non-classification

No classification is required for fertility as there were no adverse effects found. For developmentmental effects in an OECD 422 study, adverse effects were only found at maternal doses of twice the limit dose and MTD when maternal test article concentrations were not adjusted during lactation to compensate for increasing maternal food intake.

Additional information